U.S. patent number 6,835,559 [Application Number 10/240,056] was granted by the patent office on 2004-12-28 for process for the production of optically active .beta.-amino alcohols.
This patent grant is currently assigned to Daiichi Fine Chemical Co., Ltd.. Invention is credited to Michihiko Kataoka, Shinji Kita, Tadanori Morikawa, Keiji Sakamoto, Sakayu Shimizu, Kazuya Tsuzaki.
United States Patent |
6,835,559 |
Sakamoto , et al. |
December 28, 2004 |
Process for the production of optically active .beta.-amino
alcohols
Abstract
A process for producing an optical active .beta.-amino alcohol,
the method comprising the step of allowing at least one
microorganism selected from the group consisting of microorganisms
belonging to the genus Morganella and others, to act on an
enantiomeric mixture of an .alpha.-aminoketone or a salt thereof
having the general formula (I): ##STR1## to produce an optical
active .beta.-amino alcohol with the desired optical activity
having the general formula (II) described below in a high yield as
well as in a highly selective manner: ##STR2##
Inventors: |
Sakamoto; Keiji (Takaoka,
JP), Kita; Shinji (Takaoka, JP), Tsuzaki;
Kazuya (Takaoka, JP), Morikawa; Tadanori
(Takaoka, JP), Shimizu; Sakayu (Kyoto, JP),
Kataoka; Michihiko (Kyoto, JP) |
Assignee: |
Daiichi Fine Chemical Co., Ltd.
(Toyama, JP)
|
Family
ID: |
18604966 |
Appl.
No.: |
10/240,056 |
Filed: |
January 22, 2003 |
PCT
Filed: |
March 02, 2001 |
PCT No.: |
PCT/JP01/01628 |
371(c)(1),(2),(4) Date: |
January 22, 2003 |
PCT
Pub. No.: |
WO01/73100 |
PCT
Pub. Date: |
October 04, 2001 |
Foreign Application Priority Data
|
|
|
|
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Mar 28, 2000 [JP] |
|
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2000-89182 |
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Current U.S.
Class: |
435/128; 435/169;
435/822; 435/865; 435/911; 435/917; 435/918; 435/914; 435/877;
435/170; 435/280 |
Current CPC
Class: |
C12P
13/001 (20130101); C12P 41/002 (20130101); C12P
13/008 (20130101); Y10S 435/877 (20130101); Y10S
435/865 (20130101); Y10S 435/914 (20130101); Y10S
435/911 (20130101); Y10S 435/918 (20130101); Y10S
435/917 (20130101); Y10S 435/822 (20130101) |
Current International
Class: |
C12P
41/00 (20060101); C12P 13/00 (20060101); C12P
013/00 () |
Field of
Search: |
;435/128,169,170,280 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
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0654534A2 |
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May 1995 |
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EP |
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0779366 |
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Jun 1997 |
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EP |
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10-248591 |
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Sep 1998 |
|
JP |
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WO98/12155 |
|
Mar 1998 |
|
WO |
|
Other References
"Enantioselective Synthesis of Both Enantiomers of Cathinone via
the Microbiological Reduction of 2-Azido-1-Phenyl-1-Propanone" as
published in: Journal Organic Chemistry 1994, vol. 59, pp.
8288-8291 (4 pages) Authors: Pascale Besse, Henri Veschambre,
Michael Dickman and Robert Chenevert. .
"Enantioselective Synthesis of Optically Active B-Aminoalcohols via
Asymmetric Reduction" as published in: Tetrahedron Asymmetry, 1992,
vol. 3, No. 3, pp. 341-342 (2 pages) (published in Great Britain)
Authors: Byung Tae Cho and Yu Sung Chun. .
"Asymmetric Amplifying Phenomena in Enantioselective Addition of
Diethylzine to Benzaldehyde" as published in: Journal American
Chemical Society,1988, vol. 110, pp. 7877-7878 (2 pages) Authors:
N. Oguni, Y. Matsuda, and T. Kaneko. .
"Occurrence of an Inducible NADP -Dependent D-Phenylserine
Dehydrogenase in Pseudomonas Syringae NK-15 Isolated from Soil"
Authors: Kanoktip Packdibamrung, Haruo Misono, Shinji Nagata and
Susumu Nagasaki of the Laboratory of Applied Microbiology,
Department of Bioresources Science, Kochi University, Nankoku,
Kochi 783, Japan, Received May 24, 1993, 4 pages. .
"Biological Method for Preparation of Both Pure Enantiomers of a
Chiral Compound: The Case of 2-Amino-1-Phenylethanol" as published
in Chemistry Express, vol. 4, No. 9, 1989 (6 pages) Authors:
Yoshimitsu Yamazaki, Kazuya Mochizuki and Kuniaki Hosono. .
Patent Abstracts of Japan publication No. 10-248591 published Sep.
22, 1998. .
"An Enantioselective Synthesis of (IS, 2S)-Pseudoephedrine" as
published in Tetrahedron Letters 41 (2000) pp. 953-954 (2 pages)
Authors: G. Vidyasagar Reddy, G. Venkat Rao, V. Sreevani and D.S.
Iyengar. .
Patentschrift Nr. 11332filed Oct. 18, 1953, published Mar. 12, 1956
(3 pages) and Partial English Translation (1 page). .
Patentschrift Nr. 13683, filed Jan. 19, 1956, published Aug. 27,
1957 (3 pages) and partial English translation (1 page)..
|
Primary Examiner: Lilling; Herbert J.
Attorney, Agent or Firm: Osha & May L.L.P.
Claims
What is claimed is:
1. A process for producing an optical active .beta.-amino alcohol,
the process comprising allowing at least one microorganism selected
from the group consisting of microorganisms belonging to the genus
Morganella genus, the genus Microbacterium, the genus
Sphingobacterium, the genus Nocardioides, the genus Mucor, the
genus Absidia, the genus Aspergillus, the genus Penicillium, the
genus Grifola, the genus Eurotium, the genus Ganoderma, the genus
Hypocrea, the genus Helicostylum, the genus Verticillium, the genus
Fusarium, the genus Tritirachium, the genus Mortierella, the genus
Armillariella, the genus Cylindrocarpon, the genus Klebsiella, the
genus Aureobacterium, the genus Xanthomonas, the genus Pseudomonas,
the genus Mycobacterium, the genus Sporobolomyces, the genus
Sporidiobolus, the genus Amycolatopsis, the genus Coprinus, the
genus Serratia, the genus Rhodococuss and the genus Rhodotorula to
act on an enantiomeric mixture of an .alpha.-amino ketone or a salt
thereof having the general formula (I): ##STR9##
wherein X may be the same or different and represents at least one
member selected from the group consisting of a halogen atom, lower
alkyl, hydroxyl optionally protected with a protecting group, nitro
and sulfonyl; n represents an integer of from 0 to 3; R.sup.1
represents lower alkyl; R.sup.2 and R.sup.3 may be the same or
different and represent at least one member selected from the group
consisting of a hydrogen atom and lower alkyl; and "*" represents
an asymmetric carbon,
to produce an optically active .beta.-amino alcohol compound with
the desired optical activity having the general formula (II):
##STR10##
wherein X, n, R.sup.1, R.sup.2, R.sup.3 and "*" are as previously
defined.
2. The process for producing an optically active .beta.-amino
alcohol according to claim 1, wherein the microorganism is at least
one microorganism selected from the group consisting of
microorganisms belonging to Morganella morganii, Microbacterium
arborescens, Sphingobacterium multivorum, Nocardioides simplex,
Mucor ambiguus, Mucor javanicus, Mucor fragilis, Absidia
lichtheimi, Aspergillus awamori, Aspergillus niger, Aspergillus
oryzae, Aspergillus candidus, Aspergillus oryzae var. oryzae,
Aspergillus foetidus var. acidus, Penicillium oxalicum, Grifola
frondosa, Eurotium repens, Ganoderma lucidum, Hypocrea gelatinosa,
Helicostylum nigricans, Verticillium fungicola var. fungicola,
Fusarium roseum, Tritirachium, oryzae, Mortierella isabellina,
Armillariella mellea, Cylindrocarpon sclerotigenum, Klebsiella
pneumoniae, Aureobacterium esteraromaticum, Xanthomonas sp.,
Pseudomonas putida, Mycobacterium smegmatis, Mycobacterium
diernhoferi, Mycobacterium vaccae, Mycobacterium phlei,
Mycobacterium fortuitum, Mycobacterium chlorophenolicum,
Sporobolomyces salmonicolor, Sporobolomyces coralliformis,
Sporidiobolus johnsonii, Amycolatopsis alba, Amycolatopsis azurea,
Amycolatopsis coloradensis, Amycolatopsis orientalis lurida,
Amycolatopsis orientalis orientalis, Coprinus rhizophorus, Serratia
marcescens, Rhodococcus erythropolis, Rhodococcus rhodochrous and
Rhodotorula aurantiaca.
3. The process for producing an optically active .beta.-amino
alcohol according to claim 1, wherein the microorganism is at least
one microorganism selected from the group consisting of
microorganisms belonging to the genus Morganella, the genus
Microbacterium, the genus Sphingobacterium, the genus Nocardioides,
the genus Mucor, the genus Absidia, the genus Aspergillus, the
genus Penicillium, the genus Grifola, the genus Eurotium, the genus
Ganoderma, the genus Hypocrea, the genus Helicostylum, the genus
Verticillium, the genus Fusarium, the genus Tritirachium, the genus
Mortierella, the genus Armillariella, the genus Cylindrocarpon, the
genus Klebsiella, the genus Aureobacterium, the genus Xanthomonas,
the genus Pseudomonas, the genus Mycobacterium, the genus
Sporobolomyces, the genus Sporidiobolus and the Rhodococuss genus;
and the .beta.-amino alcohol having the general formula (II) is
(1S,2S)-amino alcohol.
4. The process for producing an optically active .beta.-amino
alcohol according to claim 3, wherein the microorganism is at least
one microorganism selected from the group consisting of
microorganisms belonging to Morganella morganii, Microbacterium
arborescens, Sphingobacterium multivorum, Nocardioides simplex,
Mucor ambiguus, Mucor javanicus, Mucor fragilis, Absidia
lichtheimi, Aspergillus awamori, Aspergillus niger, Aspergillus
oryzae, Aspergillus candidus, Aspergillus oryzae var. oryzae,
Aspergillus foetidus var. acidus, Penicillium oxalicum, Grifola
frondosa, Eurotium repens, Ganoderma lucidum, Hypocrea gelatinosa,
Helicostylum nigricans, Verticillium fungicola var. fungicola,
Fusarium roseum, Tritirachium, oryzae, Mortierella isabellina,
Armillariella mellea, Cylindrocarpon sclerotigenum, Klebsiella
pneumoniae, Aureobacterium esteraromaticum, Xanthomonas sp.,
Pseudomonas putida, Mycobacterium smegmatis, Mycobacterium
diernhoferi, Mycobacterium vaccae, Mycobacterium phlei,
Mycobacterium fortuitum, Mycobacterium chlorophenolicum,
Sporobolomyces salmonicolor, Sporobolomyces coralliformis,
Sporidiobolus johnsonii, Rhodococus erythropolis and Rhodococcus
rhodochrous.
5. The process for producing an optically active .beta.-amino
alcohol according to claim 1, wherein the microorganism is at least
one microorganism selected from the group consisting of
microorganisms belonging to the genus Amycolaptopsis, the genus
Coprinus, the genus Serratia, the genus Rhodococuss and the genus
Rhodotorula; and the optically active .beta.-amino alcohol having
the general formula (II) is (1R,2R)-amino alcohol.
6. The process for producing an optically active .beta.-amino
alcohol according to claim 5, wherein the microorganism is at least
one microorganism selected from the group consisting of
microorganisms belonging to Amycolatopsis alba, Amycolatopsis
azurea, Amycolatopsis coloradensis, Amycolatopsis orientalis
lurida, Amycolatopsis orientalis orientalis, Coprinus rhizophorus,
Serratia marcescens, Rhodococcus erythropolis, Rhodococcus
rhodochrous and Rhodotorula aurantiaca.
7. The process for producing an optically active .beta.-amino
alcohol according to any of claims 1-6, wherein the microorganism
is cultured in a medium to which there has been added an activity
inducer having the general formula (III): ##STR11##
wherein R.sup.4 represents lower alkyl; R.sup.5 and R.sup.6 may be
the same or different and each represents a hydrogen atom, lower
alkyl or acyl; and Y represents C.dbd.O or CH--OH.
Description
TECHNICAL FIELD
This invention relates to a process for producing optically active
.beta.-amino alcohols. More particularly, it relates to a process
for producing optically active .beta.-amino alcohols which are of
value as drugs or their intermediates.
BACKGROUND ART
Ephedrines have been used for purposes of perspiration, antipyresis
and cough soothing from the olden times, and particularly,
d-pseudoephedrine is known to possess anti-inflammatory action.
Pharmacological action such as vasoconstriction, blood pressure
elevation, or perspiration is known for 1-ephedrine and it is used
in therapy as a sympathomimetic agent. 1-Ephedrine is also used in
the treatment of bronchial asthma. Specifically, processes for the
production of optically active .beta.-amino alcohols, including
optically active ephedrines, are useful in the manufacture of drugs
and their intermediates; thus, there is a need for efficient
production processes.
In the conventional process for producing a .beta.-amino alcohol
with the desired optical activity, there was used a process by
which a racemic .beta.-amino alcohol is obtained and then a
specific optically active form is produced by optical resolution or
asymmetric synthesis among others.
However, since the racemic .beta.-amino alcohol has two asymmetric
carbons within its molecule, complicated steps had to be followed
to obtain the specific optically active form. For example,
according to Ger. (East) 13683 (Aug. 27, 1957), optically active
phenylacetylcarbinol was produced from benzaldehyde by fermentation
utilizing yeast and erythro-1-2-methylamino-1-phenyl-1-propanol
(i.e., 1-ephedrine) could be produced by reductively condensing
methylamine to the optically active phenylacetylcarbinol.
To obtain pseudoephedrine, the production is possible as described
in U.S. Pat. No. 4,237,304: an oxazoline is formed from 1-ephedrine
produced by the method described in Ger. (East) 13683 (Aug. 27,
1957), using acetic anhydride, and then the oxazoline is hydrolyzed
through inversion to the threo form (i.e., d-pseudoephedrine).
As stated above, to produce pseudoephedrine with the desired
optical activity from 2-methylamino-1-phenyl-1-propanone, steps are
necessary such that ephedrine in the optical active erythro form is
once produced and then it is inverted to the threo form. Hence,
there arise problems that the number of steps grows and leads to
complication and that the yields lower.
Furthermore, in the production of the pseudoephedrine while a
substantial amount of diastereomers is produced as byproducts
during the reduction of the starting ketone, the recovery of the
diastereomers for their use as raw material is difficult, which is
economically disadvantageous.
In addition, according to the method as described in the
publication of JP, 8-98697, A, it is possible to produce an
optically active 2-amino-1-phenylethanol derivative from a
2-amino-1-phenylethanol compound having one asymmetric carbon atom
within its molecule through the use of a specific microorganism.
The present state of art is, however, that there has been no
efficient process for producing .beta.-amino alcohol having two
asymmetric carbon atoms.
DISCLOSURE OF THE INVENTION
This invention has been made in view of the above-indicated
circumstances and it aims at producing a .beta.-amino alcohol
having the desired optical activity from an enantiomeric mixture of
an .alpha.-aminoketone compound or its salt in a high yield as well
as in a highly selective manner with a simple process while
sufficiently preventing the generation of diastereomeric
byproducts.
The present inventors repeated studies diligently to solve the
above-stated problems; consequently, it was discovered that by
utilizing specific microorganisms only one enantiomer of the
enantiomeric mixture of an .alpha.-aminoketone compound or its salt
could be reduced to produce the only desired kind among the
corresponding four kinds of .beta.-amino alcohols in a high yield
as well as in a highly selective manner. This led to the completion
of the present invention.
Specifically, the process for producing an optically active
.beta.-amino alcohol according to this invention comprises allowing
at least one microorganism selected from the group consisting of
microorganisms belonging to the genus Morganella, the genus
Microbacterium, the genus Sphingobacterium, the genus Nocardicides,
the genus Mucor, the genus Absidia, the genus Aspergillus, the
genus Penicillium, the genus Grifola, the genus Eurotium, the genus
Ganoderma, the genus Hypocrea, the genus Helicostylum, the genus
Verticillium, the genus Fusarium, the genus Tritirachium, the genus
Mortierella, the genus Armillariella, the genus Cylindrocarpon, the
genus Kiebsiella, the genus Aureobacterium, the genus Xanthomonas,
the genus Pseudomonas, the genus Mycobacterium, the genus
Sporobolomyces, the genus Sporidiobolus, the genus Amycolatopsis,
the genus Coprinus, the genus Serratia, the genus Rhodococuss and
the genus Rhodotorula to act on an enantiomeric mixture of an
.alpha.-aminoketone or a salt thereof having the general formula
(1): ##STR3##
wherein X may be the same or different and represents at least one
member selected from the group consisting of a halogen atom, lower
alkyl, hydroxyl optionally protected with a protecting group, nitro
and sulfonyl; n represents an integer of from 0 to 3; R.sup.1
represents lower alkyl; R.sup.2 and R.sup.3 may be the same or
different and represent at least one member selected from the group
consisting of a hydrogen atom and lower alkyl; and "*" represents
an asymmetric carbon, to produce an optically active .beta.-amino
alcohol compound with the desired optical activity having the
general formula (II): ##STR4##
wherein X, n, R.sup.1, R.sup.2, R.sup.3 and "*" are as previously
defined.
The microorganism according to this invention is preferably at
least one microorganism selected from the group consisting of
microorganisms belonging to Morganella morganii, Microbacterium
arborescens, Sphingobacterium multivorum, Nocardioides simplex,
Mucor ambiguus, Mucor javanicus, Mucor fragilis, Absidia
lichtheimi, Aspergillus awamori, Aspergillus niger, Aspergillus
oryzae, Aspergillus candidus, Aspergillus oryzae var. oryzae,
Aspergillus foetidus var. acidus, Penicillium oxalicum, Grifola
frondosa, Eurotium repens, Ganoderma lucidum, Hypocrea gelatinosa,
Helicostylum nigricans, Verticillium fungicola var. fungicola,
Fusarium roseum, Tritirachium oryzae, Mortierella isabellina,
Armillariella mellea, Cylindrocarpon sclerotigenum, Klebsiella
pneumoniae, Aureobacterium esteraromaticum, Xanthomonas sp.,
Pseudomonas putida, Mycobacterium smegmatis, Mycobacterium
diernhoferi, Mycobacterium vaccae, Mycobacterium phlei,
Mycobacterium fortuitum, Mycobacterium chlorophenolicum,
Sporobolomyces salmonicolor, Sporobolomyces coralliformis,
Sporidiobolus johnsonii, Amycolatopsis alba, Amycolatopsis azurea,
Amycolatopsis coloradensis, Amycolatopsis orientalis lurida,
Amycolatopsis orientalis orientalis, Coprinus rhizophorus, Serratia
marcescens, Rhodococcus erythropolis, Rhodococcus rhodochrous and
Rhodotorula aurantiaca.
In this invention the microorganism is preferably at least one
microorganism selected from the group consisting of microorganisms
belonging to the genus Morganella, the genus Microbacterium, the
genus Sphingobacterium, the genus Nocardioides, the genus Mucor,
the genus Absidia, the genus Aspergillus, the genus Penicillium,
the genus Grifola, the genus Eurotium, the genus Ganoderma, the
genus Hypocrea, the genus Helicostylum, the genus Verticillium, the
genus Fusarium, the genus Tritirachium, the genus Mortierella, the
genus Armillariella, the genus Cylindrocarpon, the genus
Klebsiella, the genus Aureobacterium, the genus Xanthomonas, the
genus Pseudomonas, the genus Mycobacterium, the genus
Sporobolomyces, the genus Sporidiobolus and the genus Rhodococuss.
More specifically, it is preferably a microorganism selected from
the group consisting of microorganisms belonging to Morganella
morganii, Microbacterium arborescens, Sphingobacterium multivorum,
Nocardioides simplex, Mucor ambiguus, Mucor javanicus, Mucor
fragilis, Absidia lichtheimi, Aspergillus awamori, Aspergillus
niger, Aspergillus oryzae, Aspergillus candidus, Aspergillus oryzae
var. oryzae, Aspergillus foetidus var. acidus, Penicillium
oxalicum, Grifola frondosa, Eurotium repens, Ganoderma lucidum,
Hypocrea gelatinosa, Helicostylum nigricans, Verticillium fungicola
var. fungicola, Fusarium roseum, Tritirachium, oryzae, Mortierella
isabellina, Armillariella mellea, Cylindrocarpon sclerotigenum,
Klebsiella pneumoniae, Aureobacterium esteraromaticum, Xanthomonas
sp., Pseudomonas putida, Mycobacterium smegmatis, Mycobacterium
diernhoferi, Mycobacterium vaccae, Mycobacterium phlei,
Mycobacterium fortuitum, Mycobacterium chlorophenolicum,
Sporobolomyces salmonicolor, Sporobolomyces coralliformis,
Sporidiobolus johnsonii, Rhodococus erythropolis and Rhodococcus
rhodochrous. By utilizing such microorganisms, (1S,2S)-amino
alcohols tend to be obtained in simple processes as the optically
active .beta.-amino alcohols represented by the general formula
(II) in high yields as well as in a highly selective manner.
Further, the microorganism is preferably at least one microorganism
selected from the group consisting of microorganisms belonging to
the genus Amycolaptopsis, the genus Coprinus, the genus Serratia,
the genus Rhodococuss and the genus Rhodotorula. More specifically,
it is preferably a microorganism selected from the group consisting
of microorganisms belonging to Amycolatopsis alba, Amycolatopsis
azurea, Amycolatopsis coloradensis, Amycolatopsis orientalis
lurida, Amycolatopsis orientalis orientalis, Coprinus rhizophorus,
Serratia marcescens, Rhodococcus erythropolis, Rhodococcus
rhodochrous and Rhodotorula aurantiaca. By utilizing such
microorganisms, (1R,2R)-amino alcohols tend to be obtained in
simple processes as the optically active .beta.-amino alcohols
represented by the general formula (II) in high yields as well as
in a highly selective manner.
Still further, in this invention the microorganism may be cultured
in a medium to which there has been added an activity inducer
having the general formula (III): ##STR5##
wherein R.sup.4 represents lower alkyl; R.sup.5 and R.sup.6 may be
the same or different and each represents a hydrogen atom, lower
alkyl or acyl; and Y represents C.dbd.O or CH--OH. The mediation of
such an activity inducer renders the production of an optically
active .beta.-amino alcohol more efficient.
BRIEF DESCRIPTION OF THE DRAWINGS
FIGS. 1A through 1D are representations showing the structures of
the optically active .beta.-amino alcohols described below,
including their absolute configurations.
FIG. 1A shows a .beta.-amino alcohol with the (1S,2S) configuration
obtained according to this invention. FIG. 1B shows
(1S,2S)-(+)-pseudoephedrine obtained according to the invention.
FIG. 1C shows a .beta.-amino alcohol with the (1R,2R) configuration
obtained according to the invention. FIG. 1D shows
(1R,2R)-(-)-pseudoephedrine obtained according to this
invention.
BEST MODE FOR CARRYING OUT THE INVENTION
The preferred embodiments of this invention will be described in
detail hereafter.
The process for producing an optically active .beta.-amino alcohol
according to this invention comprises allowing at least one
microorganism selected from the group consisting of microorganisms
belonging to the genus Morganella, the genus Microbacterium, the
genus Sphingobacterium, the genus Nocardioides, the genus Mucor,
the genus Absidia, the genus Aspergillus, the genus Penicillium,
the genus Grifola, the genus Eurotium, the genus Ganoderma, the
genus Hypocrea, the genus Helicostylum, the genus Verticillium, the
genus Fusarium, the genus Tritirachium, the genus Mortierella, the
genus Armillanella, the genus Cylindrocarpon, the genus Kiebsiella,
the genus Aureobacterium, the genus Xanthomonas, the genus
Pseudomonas, the genus Mycobacterium, the genus Sporobolomyces, the
genus Sporidiobolus, the genus Amycolatopsis, the genus Coprinus,
the genus Serratia, the genus Rhodococuss, and the genus
Rhodotorula to act on an enantiomeric mixture of an .alpha.-amino
ketone compound or a salt thereof having the general formula (I):
##STR6##
wherein X may be the same or different and represents at least one
member selected from the group consisting of a halogen atom, lower
alkyl, hydroxyl optionally protected with a protecting group, nitro
and sulfonyl; n represents an integer of from 0 to 3; R.sup.1
represents lower alkyl; R.sup.2 and R.sup.3 may be the same or
different and represent at least one member selected from the group
consisting of a hydrogen atom and lower alkyl; and "*" represents
an asymmetric carbon, to produce an optically active .beta.-amino
alcohol compound with the desired optical activity having the
general formula (II): ##STR7##
wherein X, n, R.sup.1, R.sup.2, R.sup.3 and "*" are as previously
defined.
The starting material used in the process for producing an
optically active .beta.-amino alcohol according to this invention
is an enantiomeric mixture of an .alpha.-aminoketone compound or a
salt thereof having the general formula (I) wherein X may be the
same or different and represents at least one member selected from
the group consisting of a halogen atom, lower alkyl, hydroxyl
optionally protected with a protecting group, nitro and sulfonyl; n
represents an integer of from 0 to 3; R.sup.1 represents lower
alkyl; R.sup.2 and R.sup.3 may be the same or different and
represent at least one member selected from the group consisting of
a hydrogen atom and lower alkyl; and "*" represents an asymmetric
carbon.
The substituent group X contained in the .alpha.-amino ketone will
be described in the following: the halogen atoms include a fluorine
atom, a chlorine atom, a bromine atom and an iodine atom.
The lower alkyl groups are preferably alkyls of from one to six
carbons and include methyl, ethyl, propyl, isopropyl, butyl,
isobutyl, s-butyl, t-butyl, pentyl, isopentyl, hexyl, and the like.
These may adopt either of straight chain and branched structures
and may have as a substituent, a halogen atom such as fluorine or
chlorine, hydroxyl, alkyl, amino, or alkoxy.
For the protecting group of the hydroxyl optionally protected with
a protecting group, there are mentioned, among others, the one that
can be removed upon treatment with water, the one that can be
removed by hydrogenation, the one that can be removed by a Lewis
acid catalyst or thiourea. The protecting groups include acyl
optionally having a substituent, silyl optionally having a
substituent, alkoxyalkyl, lower alkyl optionally having a
substituent, benzyl, p-methoxybenzyl,
2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, trityl and the
like.
The acyl groups include acetyl, chloroacetyl, dichloroacetyl,
pivaloyl, benzoyl, p-nitrobenzoyl, and the like; they may also have
a substituent such as hydroxyl, alkyl, alkoxy, nitro, a halogen
atom, or the like. The silyl groups include trimethylsilyl,
t-butyldimethylsilyl, triarylsilyl, and the like; they may also
have a substituent such as alkyl, aryl, hydroxyl, alkoxy, nitro, a
halogen atom, or the like. The alkyl groups include methoxymethyl,
2-methoxyethoxymethyl and the like. The lower alkyl groups include
alkyls of from one to six carbons: there are mentioned methyl,
ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl,
pentyl, isopentyl, hexyl, and the like. These may adopt either of
straight chain and branched structures and may have a substituent
such as a halogen atom (including fluorine and chlorine), hydroxyl,
alkyl, amino, or alkoxy.
The X may be nitro or sulfonyl, and specifically, methylsulfonyl is
mentioned among others.
In addition, the number n of X is an integer of from 0 to 3,
preferably 0.
R.sup.1 in the general formula (I) represents lower alkyl. Such
lower alkyls are preferably alkyls of from one to six carbons:
there are mentioned methyl, ethyl, propyl, isopropyl, butyl,
isobutyl, s-butyl, t-butyl, pentyl, isopentyl, hexyl, and the like.
These may adopt either of straight chain and branched
structures.
R.sup.2 and R.sup.3 represent a hydrogen atom or lower alkyl. The
lower alkyls include alkyls of from one to six carbons: there are
mentioned methyl, ethyl, propyl, isopropyl, butyl, isobutyl,
s-butyl, t-butyl, pentyl, isopentyl, hexyl, and the like. These may
adopt either of straight chain and branched structures.
The salts of the .alpha.-aminoketone compound include the salts of
inorganic acids such as hydrochloride, sulfate, nitrate, phosphate,
and carbonate and the salts of organic acids such as acetate and
citrate.
The .alpha.-aminoketone can readily be synthesized by halogenating
the .alpha.-carbon of the corresponding 1-phenyl ketone derivative
(e.g., bromination) and substituting the halogen such as bromo for
amine (Ger. (East) 11, 332, Mar. 12, 1956).
The microorganism according to this invention is that which act on
the enantiomeric mixture of the a .alpha.-minoketone having the
general formula (I) or a salt thereof. Such microorganism is one
selected from the group consisting of microorganisms belonging to
the genus Morganella, the genus Microbacterium, the genus
Sphingobacterium, the genus Nocardioides, the genus Mucor, the
genus Absidia, the genus Aspergillus, the genus Penicillium, the
genus Grifola, the genus Eurotium, the genus Ganoderma, the genus
Hypocrea, the genus Helicostylum, the genus Verticillium, the genus
Fusarium, the genus Tritirachium, the genus Mortierella, the genus
Armillariella, the genus Cylindrocarpon, the genus Klebsiella, the
genus Aureobacterium, the genus Xanthomonas, the genus Pseudomonas,
the genus Mycobacterium, the genus Sporobolomyces, the genus
Sporidiobolus, the genus Amycolatopsis, the genus Coprinus, the
genus Serratia, the genus Rhodococuss and the genus Rhodotorula.
Specifically, the preferred ones include Morganella morganii IFO
3848, Microbacterium arborescens IFO 3750, Sphingobacterium
multivorum IFO 14983, Nocardioides simplex IFO 12069, Mucor
ambiguus IFO 6742, Mucor javanicus IFO 4570, Mucor fragilis IFO
6449, Absidia lichtheimi IFO 4009, Aspergillus awamori IFO 4033,
Aspergillus niger IFO 4416, Aspergillus oryzae IFO 4177,
Aspergillus oryzae IAM 2630, Aspergillus candidus IFO 5468,
Aspergillus oryzae var. oryzae IFO 6215, Aspergillus foetidus var.
acidus IFO 4121, Penicillium oxalicum IFO 5748, Grifola frondosa
IFO 30522, Eurotium repens IFO 4884, Ganoderma lucidum IFO 8346,
Hypocrea gelatinosa IFO 9165, Helicostylum nigricans IFO 8091,
Verticillium fungicola var. fungicola IFO 6624, Fusarium roseum IFO
7189, Tritirachium, oryzae IFO 7544, Mortierella isabellina IFO
8308, Armillariella mellea IFO 31616, Cylindrocarpon sclerotigenum
IFO 31855, Klebsiella pneumoniae IFO 3319, Aureobacterium
esteraromaticum IFO 3751, Xanthomonas sp. IFO 3084, Pseudomonas
putida IFO 14796, Mycobacterium smegmatis IAM 12065, Mycobacterium
diernhoferi, IFO 14797, Mycobacterium vaccae IFO 14118,
Mycobacterium phlei IFO 13160, Mycobacterium fortuitum IFO 13159,
Mycobacterium chlorophenolicum IFO 15527, Sporobolomyces
salmonicolor IFO 1038, Sporobolomyces coralliformis IFO 1032,
Sporidiobolus johnsonii IFO 6903, Amycolatopsis alba IFO 15602,
Amycolatopsis azurea IFO 14573, Amycolatopsis coloradensis IFO
15804, Amycolatopsis orientalis lurida IFO 14500, Amycolatopsis
orientalis orientalis IFO 12360, IFO 12362, IFO 12806, Coprinus
rhizophorus IFO 30197, Serratia marcescens IFO 3736, Rhodococcus
erythropolis IFO 12540, Rhodococcus erythropolis MAK-34,
Rhodococcus rhodochrous IFO 15564, Rhodococcus rhodochrous IAM
12126, Rhodotorula aurantiaca IFO 0951, and the like.
Such microorganisms according to this invention permit the
production of the corresponding optically active .beta.-amino
alcohol compounds having the general formula (II), said compound
possessing the desired optical activity.
In the general formula (II), X, n, R.sup.1, R.sup.2, R.sup.3
and*are the same as those in the general formula (I). Further, the
.beta.-amino alcohols having the desired optical activity include
(1S,2S)-amino alcohol, (1S,2R)-amino alcohol, (1R,2S)-amino alcohol
and (1R,2R)-amino alcohol.
In this invention the microorganism is preferably at least one
selected from the group consisting of microorganisms belonging to
the genus Morganella genus, the genus Microbacterium, the genus
Sphingobacterium, the genus Nocardioides, the genus Mucor, the
genus Absidia, the genus Aspergillus, the genus Penicillium, the
genus Grifola, the genus Eurotium, the genus Ganoderma, the genus
Hypocrea, the genus Helicostylum, the genus Verticillium, the genus
Fusarium, the genus Tritirachium, the genus Mortierella, the genus
Armillariella, the genus Cylindrocarpon, the genus Klebsiella, the
genus Aureobacterium, the genus Xanthomonas, the genus Pseudomonas,
the genus Mycobacterium, the genus Sporobolomyces, the genus
Sporidiobolus and the genus Rhodococuss. More specifically,
preferred is a microorganism selected from the group consisting of
microorganisms belonging to Morganella morganii, Microbacterium
arborescens, Sphingobacterium multivorum, Nocardioides simplex,
Mucor ambiguus, Mucor javanicus, Mucor fragilis, Absidia
lichtheimi, Aspergillus awamori, Aspergillus niger, Aspergillus
oryzae, Aspergillus candidus, Aspergillus oryzae var. oryzae,
Aspergillus foetidus var. acidus, Penicillium oxalicum, Grifola
frondosa, Eurotium repens, Ganoderma lucidum, Hypocrea gelatinosa,
Helicostylum nigricans, Verticillium fungicola var. fungicola,
Fusarium roseum, Tritirachium, oryzae, Mortierella isabellina,
Armillariella mellea, Cylindrocarpon sclerotigenum, Klebsiella
pneumoniae, Aureobacterium esteraromaticum, Xanthomonas sp.,
Pseudomonas putida, Mycobacterium smegmatis, Mycobacterium
diernhoferi, Mycobacterium vaccae, Mycobacterium phlei,
Mycobacterium fortuitum, Mycobacterium chlorophenolicum,
Sporobolomyces salmonicolor, Sporobolomyces coralliformis,
Sporidiobolus johnsonii, Rhodococcus erythropolis and Rhodococcus
rhodochrous. By utilizing such microorganisms, (1S,2S)-amino
alcohols tend to be obtained in simple processes as the optically
active .beta.-amino alcohols represented by the general formula
(II) in high yields as well as in a highly selective manner.
Furthermore, in this invention the microorganism is preferably at
least one selected from the group consisting of microorganisms
belonging to the genus Amycolatopsis, the genus Coprinus, the genus
Serratia, the genus Rhodococuss and the genus Rhodotorula. More
specifically, more preferred is a microorganism selected from the
group consisting of microorganisms belonging to Amycolatopsis alba,
Amycolatopsis azurea, Amycolatopsis coloradensis, Amycolatopsis
orientalis lurida, Amycolatopsis orientalis orientalis, Coprinus
rhizophorus, Serratia marcescens, Rhodococcus erythropolis,
Rhodococcus rhodochrous and Rhodotorula aurantiaca. By utilizing
such microorganisms, (1R,2R)-amino alcohols tend to be obtained in
simple processes as the optically active .beta.-amino alcohols
represented by the general formula (II) in high yields as well as
in a highly selective manner.
In addition, the microorganisms according to this invention include
(1S,2S)-amino alcohol producing bacteria that selectively produce
(1S,2S) forms among the optically active .beta.-amino alcohol
compounds and (1R,2R)-amino alcohol producing bacteria that
selectively produce (1R,2R) forms among the optically active
.beta.-amino alcohol compounds.
By allowing the action of the (1S,2S)-amino alcohol producing
bacteria, there can be obtained, for example,
d-threo-2-methylamino-1-phenylpropanol(d-pseudoephedrine),
d-threo-2-dimethylamino-1-phenylpropanol(d-methylpseudoephedrine),
(1S,2S)-.alpha.-(1-aminoethyl)-benzylalcohol(d-norpseudoephedrine),
(1S,2S)-1-(p-hydroxyphenyl)-2-methylamino-1-propanol,
(1S,2S)-.alpha.-(1-aminoethyl)-2,5-dimethoxy-benzylalcohol,
(1S,2S)-1-(m-hydroxyphenyl)-2-amino-1-propanol,
(1S,2S)-1-(p-hydroxyphenyl)-2-amino-1-propanol,
(1S,2S)-1-phenyl-2-ethylamino-1-propanol,
(1S,2S)-1-phenyl-2-amino-1-butanol and
(1S,2S)-1-phenyl-2-methylamino-1-butanol. By allowing the action of
the (1R,2R)-amino alcohol producing bacteria, there can be
obtained, for example,
1-threo-2-methylamino-1-phenylpropanol(1-pseudoephedrine),
1-threo-2-dimethylamino-1-phenylpropanol(1-methylpseudoephedrine),
(1R,2R)-.alpha.-(1-aminoethyl)-benzylalcohol(1-norpseudoephedrine),
(1R,2R)-1-(p-hydroxyphenyl)-2-methylamino-1-propanol,
(1R,2R)-.alpha.-(1-aminoethyl)-2,5dimethoxy-benzylalcohol,
(1R,2R)-1-(m-hydroxyphenyl)-2-amino-1-propanol,
(1R,2R)-1-(p-hydroxyphenyl)-2-amino-1-propanol,
(1R,2R)-1-phenyl-2-ethylamino-1-propanol,
(1R,2R)-1-phenyl-2-amino-1-butanol and
(1R,2R)-1-phenyl-2-methylamino-1-butanol.
Additionally, the obtained
(1S,2S)-1-(m-hydroxyphenyl)-2-amino-1-propanol can be inverted to
produce (1R,2S)-1-(m-hydroxyphenyl)-2-amino-1-propanol
(metaraminol).
Among the microorganisms according to this invention, those to
which IFO accession numbers have been designated are described in
the "List of Cultures, 10th Edition (1966)" published by Institute
for Fermentation (IFO) (non-profit organization) and are available
from the IFO. The microorganisms to which IAM accession numbers
have been designated are described in the "Catalogue of Strains,
1993" published by Institute of Molecular and Cellular Biosciences,
the Cell & Functional Polymer General Center, University of
Tokyo and are available from its preservation facilities. Further,
Rhodococcus erythropolis MAK-34 is a novel microorganism isolated
from the nature and has been deposited with National Institute of
Bioscience and Human-Technology, National Institute of Advanced
Science and Technology, METI locating at 1-3, Higashi 1-Chome,
Tsukuba, Ibaraki, JAPAN (postal code: 305-8566) as FERM BP-7451
(the date of original deposit: Feb. 15, 2001).
For the microorganism used in this invention, there can be used any
of wild-type strains, mutant strains and recombinant strains
derived by the techniques of cell engineering such as cell fusion
or by the techniques of genetic engineering such as gene
manipulations insofar as it is a microorganism capable of acting on
the enantiomeric mixture of a .alpha.-aminoketone compound of the
general formula (I) and producing the corresponding optically
active .beta.-amino alcohol of the general formula (II).
There are no particular limitations to the different conditions in
the culturing of the microorganisms, and the methods that are
ordinarily used may be carried out, where bacteria, fungi, and
yeast are cultured in suitable media, respectively. Normally, there
may be used liquid media containing carbon sources, nitrogen
sources and other nutrients. Any sources may be used for the carbon
source of the medium as long as the microorganisms can utilize
them. Specifically, there may be used sugars such as glucose,
fructose, sucrose, dextrin, starch, and sorbitol; alcohols such as
methanol, ethanol, and glycerol; organic acids such as fumaric
acid, citric acid, acetic acid, and propionic acid and their salts;
hydrocarbons such as paraffin; and mixtures of the foregoing. Any
sources may be used for the nitrogen source of the medium as long
as the microorganisms can utilize them. Specifically, there may be
used the ammonium salts of inorganic acids such as ammonium
chloride, ammonium sulfate, and ammonium phosphate; the ammonium
salts of organic acids such as ammonium fumarate and ammonium
citrate; the salts of nitric acid such as sodium nitrate and
potassium nitrate; nitrogen-containing inorganic or organic
compounds such as beef extract, yeast extract, malt extract, and
peptone; and mixtures of the foregoing. Nutrition sources may also
be added appropriately to the medium, which are used in the normal
culturing, including inorganic salts, the salts of minute metals,
and vitamins. There may also be added to the medium, a substance
for inducing the activity of a microorganism, a buffer substance
effective to maintain pH, or the like.
The substances for inducing the activity of a microorganism include
an activity inducer having the general formula (III): ##STR8##
wherein R.sup.4 represents lower alkyl; R.sup.5 and R.sup.6 may be
the same or different and each represents a hydrogen atom, lower
alkyl, or acyl; Y represents C.dbd.O or CH--OH.
The lower alkyl and acyl groups include the ones previously defined
respectively. Specifically, the preferred activity inducers include
1-amino-2-propanol, 1-amino-2-hydroxybutane,
1-acetylamino-2-propanol, 1-methylamino-2-propanol,
1-amino-2-oxopropane, 2-amino-3-hydroxybutane, and the like. When
asymmetric carbons are present in these compounds, the compounds
may be either of optically active forms and racemates, and may
appropriately be selected. The addition of these activity inducers
to medium induces the activity of microorganisms and the subsequent
generation of optically active .beta.-amino alcohols to progress
with higher efficiency as compared to the case with no such
addition. The activity inducers may be used individually, or may be
used as a mixture of plural inducers. The addition levels of such
activity inducers are desirably 0.01 to 10 wt. % relative to
medium.
Culturing of microorganisms can be carried out under the conditions
suited for their growth. Specifically, it can be done at the pH of
medium being 3-10, preferably pH 4-9 and at a temperature of
0-50.degree. C., preferably 20-40.degree. C. The culturing of
microorganisms can be carried out under aerobatic conditions or
anaerobatic conditions. The culturing time is preferably from 10 to
150 hours and should be appropriately determined for the respective
microorganisms.
The reaction method in the production of .beta.-amino alcohols
according to this invention is not particularly limited insofar as
it is a method by which the microorganism acts on the enantiomeric
mixture of the .alpha.-amino ketone compound having the general
formula (I) or a mixture thereof to produce the corresponding
optically active .beta.-amino alcohol compound having the general
formula (II). The reaction is allowed to start by mixing bacterial
cells washed with buffer or water to a aqueous solution of the
starting .alpha.-aminoketone.
The reaction conditions can be selected from the range within which
the generation of the optically active .beta.-amino alcohol
compound having the general formula (II) is not impaired. The
quantity of bacterial cell is preferably 1/100 to 1000 times, and
more preferably 1/10 to 100 times that of racemic aminoketone. The
concentration of the racemic aminoketone that is a substrate is
preferably from 0.01 to 20%, and more preferably from 0.1 to 10%.
Further, the pH of reaction solution is preferably from 5 to 9, and
more preferably from 6 to 8; the reaction temperature is preferably
from 10 to 50.degree. C., and more preferably from 20 to 40.degree.
C. Still further, the reaction time is preferably from 5 to 150
hours and it should be appropriately determined for the respective
microorganisms.
In order for the reaction to progress more efficiently, sugars
(e.g., glucose), organic acids (e.g., acetic acid), and energy
substances (e.g., glycerol) may be added. These may be respectively
used alone or may be used as a mixture thereof. The level of
addition is preferably 1/100 to 10 times that of substrate.
Coenzymes or the like may also be added. Coenzymes such as
nicotinamide adenine dinucleotide (NAD), reduced nicotinamide
adenine dinucleotide (NADH), nicotinamide adenine dinucleotide
phosphate (NADP), and reduced nicotinamide adenine dinucleotide
phosphate (NADPH) can be used alone or as a mixture of the
foregoing. The level of addition is preferably from 1/1000 to 1/5
times that of the racemic aminoketone. In addition to these
coenzymes coenzyme-regenerating enzymes such as glucose
dehydrogenase may also be added; and the level of addition is
preferably from 1/100 to 10 times that of the racemic aminoketone.
Further, sugars (e.g., glucose), organic acids (e.g., acetic acid),
and energy substances (e.g., glycerol), coenzymes,
coenzyme-regenerating enzymes, and substrates for the
coenzyme-regenerating enzymes may respectively be combined for use.
These substances are naturally accumulated in bacterial cells, but
where their addition as required can increase the reaction rate and
yield, they may be appropriately selected.
Furthermore, when a certain salt is added such that the reaction
solution may be as described above and the reaction solution is
allowed to react under that condition, the racemization of the
unreacted .alpha.-aminoketone isomer can be accelerated and its
conversion to the enantiomer that serves as the substrate for
microorganism can be progressed more efficiently. This tends to
produce the objective amino alcohol in a high yield of 50% or more
from the starting material.
The salts for accelerating the racemization of the unreacted
.alpha.-aminoketones may be the salts of weak acids such as
acetate, tartrate, benzoate, citrate, malonate, phosphate,
carbonate, p-nitrophenolate, sulfite, and borate. Preferably, there
are used phosphates (e.g., sodium dihydrogenphosphate, potassium
dihydrogenphosphate, and ammonium dihydrogenphosphate), carbonates
(e.g., sodium carbonate, sodium hydrogencarbonate, potassium
carbonate, and ammonium carbonate), citrates (e.g., sodium citrate,
potassium citrate, and ammonium citrate), for example. These
mixtures may also be used, and desirably, buffers with pH 6.0-8.0
are added to give final concentrations of from 0.01 to 1 M. For
example, in the case of a phosphate, sodium dihydrogenphosphate and
sodium monohydrogenphosphate are desirably mixed at a ratio of from
9:1 to 5:95.
The optically active .alpha.-amino alcohols produced by reaction
may be purified by conventional separation/purification means. For
example, directly from the reaction solution or after the bacterial
cells are separated, optically active .beta.-amino alcohols can be
obtained by being subjected to normal purification methods such as
membrane separation or extraction with an organic solvent (e.g.,
toluene and chloroform), column chromatography, concentration at
reduced pressure, distillation, recrystallization, and
crystallization.
The optical activity of the optically active .beta.-amino alcohol
thus produced can be determined by high performance liquid
chromatography (HPLC).
EXAMPLES
This invention will be described concretely by way of examples;
however, the scope of the invention is not to be limited by these
examples.
Preparation Example 1
Preparation of dl-2-methylamino-1-phenyl-1-propanone
Bromine (51.6 ml) was added dropwise to a mixture of
1-phenyl-1-propanone (134 g), sodium carbonate (42 g) and water
(200 ml), and reaction was allowed to take place at 70.degree. C.
for 3 hours to give a reaction mixture. To the reaction mixture was
added 40% aqueous monomethylamine solution (350 ml). After allowing
to react at 40.degree. C. for 1 hour, the reaction product was
extracted into chloroform (1 l ). The reaction product in the
chloroform layer was then extracted with dilute hydrochloric acid
(100 ml), and activated carbon (3 g) was added to the aqueous layer
and filtrated. The filtrate was concentrated to give
dl-2-methylamino-1-phenyl-1-propanone hydrochloride (89 g).
Example 1
Production of d-(1S,2S)-pseudoephedrine
Microbacterium arborescens IFO 3750 was inoculated to a medium (5
ml) containing 1% glucose, 0.5% peptone, and 0.3% yeast extract,
and shake-culturing was carried out at 30.degree. C. for 48 hours.
After the cultured solution was centrifuged to give bacterial
cells, the cells were placed into a test tube. To this was added
0.1 M sodium phosphate buffer (pH 7.0, 1 ml) and suspended. To this
was added dl-2-methylamino-1-phenyl-1-propanone hydrochloride (1
mg) and reaction was allowed to take place under shaking at
30.degree. C. for 24 hours. After the reaction, the reaction
solution was centrifuged to remove the bacterial cells and the
supernatant was subjected to HPLC to give optically active
pseudoephedrine: .mu. Bondapakphenyl manufactured by Waters Inc.;
diameter of 4 mm; length of 300 mm; eluent-0.05 M sodium phosphate
buffer (containing 7% acetonitrile); pH 5.0; flow rate of 0.8
ml/min; and detection light wavelength at 220 nm.
The absolute configuration and optical purity were determined with
HPLC (Column Sumichiral AGP manufactured by Sumika Chemical
Analysis Service; diameter of 4 mm; length of 150 mm; 0.03 M sodium
phosphate buffer; pH 7.0; flow rate of 0.5 ml/min; and detection
light wavelength at 220 nm). Consequently, only d-pseudoephedrine
was obtained selectively, as shown in Table 1.
The produced amounts will be all shown in terms of the amounts of
converted hydrochloride hereafter.
Examples 2 to 12
Production of d-(1S,2S)-pseudoephedrine
Except that the microorganisms shown in Table 1 were used in place
of Microbacterium arborescens IFO 3750, optically active
pseudoephedrine was obtained similarly to Example 1. Consequently,
only d-pseudoephedrine was obtained selectively, as shown in Table
1.
TABLE 1 Optical activity (%) Produced Example Microorganism
d-pseudo l-pseudo amount No. genus IFO d-ephedrine l-ephedrine
ephedrine ephedrine (mg/mL) Example 1 Microbacterium 3750 0 0 100 0
0.16 arborescens Example 2 Klebsiella 3319 0 0 100 0 0.3 pneumoniae
Example 3 Aureobacterium 3751 0 0 100 0 0.14 esteraromaticum
Example 4 Xanthomonas sp. 3084 0 0 100 0 0.049 Example 5
Pseudomonas 14796 0 0 100 0 0.1 putida Example 6 Mycobacterium IAM
0 0 100 0 0.24 smegmatis 12065 Example 7 Mycobacterum 14797 0 0 100
0 0.25 diernhoferi Example 8 Mycobacterum 14118 0 0 100 0 0.28
vaccae Example 9 Mortierella 8308 0 0 100 0 0.15 isabellina Example
10 Cyllindrocarpon 31855 0 0 100 0 0.09 sclerotigenum Example 11
Sporidiobolus 6903 0 0 100 0 0.07 johnsonii Example 12 Rhodococcus
MAK-34 0 0 100 0 0.3 erythropolis
Examples 13 to 37
Production of d-(1S,2S)-pseudoephedrine
Except that the microorganisms shown in Table 2 were used in place
of Microbacterium arborescens IFO 3750, optically active
pseudoephedrine was obtained similarly to Example 1. The produced
amounts and optical purities of pseudoephedrine are shown In Table
2.
TABLE 2 Microorganism produced amount optical purity (%) Example
No. genus IFO (mg/ml) d-pseudoephedrine Example 13 Nocardioides
simplex 12069 0.35 99 Example 14 Mycobacterium phlei 13160 0.27
95.6 Example 15 Mucor ambiguus 6742 0.07 93 Example 16 Mucor
javanicus 4570 0.04 95 Example 17 Mucor fragilis 6449 0.17 90
Example 18 Absidia lichtheimi 4009 0.04 93 Example 19 Aspergillus
awamori 4033 0.18 93 Example 20 Aspergillus niger 4416 0.11 90
Example 21 Aspergillus oryzae 4177 0.18 91 Example 22 Aspergillus
candidus 5468 0.07 94 Example 23 Aspergillus oryzae IAM2630 0.08 92
Example 24 Aspergillus oryzae 6215 0.05 95 var. oryzae Example 25
Penicillium oxalicum 5748 0.06 94 Example 26 Grifola frondosa 30522
0.08 92 Example 27 Eurotium repens 4884 0.08 92 Example 28
Ganoderma lucidum 8346 0.05 92.2 Example 29 Hypocrea gelatinosa
9165 0.27 92.2 Example 30 Helicostylum 8091 0.27 93.2 nigricans
Example 31 Aspergillus foetidus 4121 0.43 91.9 var. acidus Example
32 Verticillium fungicola 6624 0.10 92.7 var. fungicola Example 33
Fusarium roseum 7189 0.40 89.6 Example 34 Tritirachium oryzae 7544
0.34 92 Example 35 Armillariella mellea 31616 0.28 91 Example 36
Sporobolomyces 1038 0.14 95 salmonicolor Example 37 Sporobolomyces
1032 0.2 95 coralliformis
Example 38
Production of d-(1S,2S)-pseudoephedrine
Morganella morganii IFO 3848 was inoculated to a medium containing
1% glucose, 0.5% peptone, and 0.3% yeast extract, and
shake-culturing was aerobically carried out at 30.degree. C. for 48
hours. After this cultured solution (5 ml) was centrifuged to give
bacterial cells, the cells were dried in the air and the resulting
dried bacterial cells were suspended in 1 ml of 0.05 M Tris
hydrochloric acid buffer (pH 7.5). To the aforementioned dried
bacterial cell suspension were added glucose (50 mg), glucose
dehydrogenase (0.2 mg), NADP (0.6 mg), NAD (0.6 mg), and
dl-2-methylamino-1-phenyl-1-propanone hydrochloride (10 mg) and
reciprocation-shaking was carried out at 28.degree. C. and at 300
rpm. After allowing to react for 48 hours, the reaction solution
was measured for the produced amount and the optical activity of
pseudoephedrine by HPLC similarly to Example 1. Consequently, only
d-pseudoephedrine was obtained selectively, as shown in Table
3.
TABLE 3 Optical purity (%) Produced Microorganism d-pseudo-
l-pseudo- amount Example No. Genus IFO d-ephedrine l-ephedrine
ephedrine ephedrine (mg/ml) Example 38 Morganella morganii 3848 0 0
100 0 0.79
Example 39
Production of d-(1S,2S)-pseudoephedrine hydrochloride
Mycobacterium smegmatis IAM-12065 was inoculated to a medium
containing 1% glucose, 0.5% peptone, and 0.3% yeast extract, and
shake-culturing was aerobically carried out at 30.degree. C. for 48
hours. After the cultured solution (1 l) was centrifuged to give
bacterial cells, the cells was suspended in 50 ml of water, and
after addition of dl-2-methylamino-1-phenyl-1-propanone
hydrochloride (0.5 g), reciprocation-shaking was carried out at
30.degree. C. and at 150 rpm. One hundred hours after the start of
shaking, 7.0 g/l of d-pseudoephedrine was produced in the reaction
solution. After the reaction solution was centrifuged to remove the
bacterial cells, the pH was adjusted to 12 or greater by addition
of sodium hydroxide. Methylene chloride (100 ml) was added to this
reaction solution and the reaction product was extracted. The
solvent was removed, hydrochloric acid was added, and then
concentration to dryness yielded a hydrochloride salt. The
hydrochloride salt was dissolved by addition of ethanol and further
addition of ether crystallized the reaction product. Consequently,
d-pseudoephedrine hydrochloride was obtained. The resulting
d-pseudoephedrine crystals (0.32 g) were analyzed on HPLC (Column
Sumichiral AGP manufactured by Sumika Chemical Analysis Service;
diameter of 4 mm; length of 150 mm; 0.03 M sodium phosphate buffer;
pH 7.0; flow rate of 0.5 ml/min; detection light wavelength at UV
220 nm) and the optical activity was found to be 100%.
Example 40
Production of d-(1S,2S)-methylpseudoephedrine
Mycobacterium smegmatis IAM-12065 was inoculated to a medium
containing 1% glucose, 0.5% peptone, and 0.3% yeast extract, and
shake-culturing was aerobically carried out at 30.degree. C. for 48
hours. The cultured solution (1 l) was filtrated to give bacterial
cells, the resulting cells were washed with water, and water was
added to form 50 ml of suspension. To the suspension was added 100
mg of 2-dimethylamino-1-phenyl-1-propanone hydrochloride (2 g/l),
and reciprocation-shaking was carried out at 30.degree. C. and at
150 rpm for 48 hours. When the reaction solution was analyzed on
HPLC (Column Sumichiral AGP manufactured by Sumika Chemical
Analytical Center; diameter of 4 mm; length of 150 mm; 0.03 M
sodium phosphate buffer; pH 7.0; flow rate of 0.5 ml/min; detection
light wavelength at UV 220 nm), it was found that
d-(1S,2S)-methylpseudoephedrine was produced at 0.23 g/l and its
optical activity was 77%.
Example 41
Production of l-(1R,2R)-pseudoephedrine
Amycolatopsis alba IFO 15602 was inoculated to a medium containing
1% glucose, 0.5% peptone and 0.3% yeast extract, and
shake-culturing was aerobically carried out at 30.degree. C. for 48
hours. The cultured solution (5 ml) was centrifuged to give
bacterial cells. After the cells were suspended in 1 ml of 0.1 M
sodium phosphate (pH 7.0) and dl-2-methylamino-1-phenyl-1-propanone
hydrochloride (1 mg) was added thereto, reaction was allowed to
take place by carrying out reciprocation-shaking at 30.degree. C.
and at 150 rpm for 48 hours. When the reaction solution was
analyzed on HPLC (Column Sumichiral AGP manufactured by Sumitomo
Chemical Analytical Center Co. Ltd.; diameter of 4 mm; length of
150 mm; 0.03 M sodium phosphate buffer; pH 7.0; flow rate of 0.5
ml/min; detection light wavelength at UV 220 nm), it was found that
1-pseudoephedrine was produced selectively. The result is shown in
Table 4.
Examples 42 to 46
Production of l-(1R,2R)-pseudoephedrine
Except that the microorganisms shown in Table 4 were used in place
of Amycolatopsis alba IFO 15602, optically active pseudoephedrine
was obtained similarly to Example 41. Consequently, only
l-pseudoephedrine was obtained selectively, as shown in Table
4.
TABLE 4 Optical activity (%) Produced Microorganism d- l- d-pseudo
l-pseudo amount Example No. genus IFO ephedrine ephedrine ephedrine
ephedrine (mg/ml) Example 41 Amycolatopsis alba 15602 0 0 0 100
0.33 Example 42 Amycolatopsis 14573 0 0 0 100 0.064 azurea Example
43 Amycolatopsis 15804 0 0 0 100 0.5 coloradensis Example 44
Amycolatopsis 14500 0 0 0 100 0.18 orientalis lurida Example 45
Amycolatopsis 12360 0 0 0 100 0.5 orientalis orientalis Example 46
Serratia marcescens 3736 0 0 0 100 0.47
Examples 47 and 48
Production of l-(1R,2R)-pseudoephedrine
Except that the microorganisms shown in Table 5 were used in place
of Amycolatopsis alba IFO 15602, optically active pseudoephedrine
was obtained similarly to Example 41. The produced amounts and
optical purities of 1-pseudoephedrine are shown in Table 5.
TABLE 5 Produced Microorganism Optical purity (%) amount Example
No. genus IFO 1-pseudoephedrine (mg/ml) Example 47 Rhodococcus
12540 98.6 0.11 erythropolis Example 48 Rhodococcus 15564 96.6 0.10
rhodochrous
Example 49
Production of l-(1R,2R)-pseudoephedrine
Coprinus rhizophorus IFO 30197 was inoculated to a medium
containing 1% glucose, 0.5% peptone and 0.3% yeast extract, and
culturing was aerobically carried out at 30.degree. C. for 48
hours. After this cultured solution (5 ml) was centrifuged to give
bacterial cells, the cells were dried in the air and the resulting
dried bacterial cells were suspended in 1 ml of 0.05 M Tris
hydrochloric acid buffer (pH 7.5). To this were added glucose (50
mg), glucose dehydrogenase (0.2 mg), NADP (0.6 mg), NAD (0.6 mg)
and dl-2-methylamino-1-phenyl-1-propanone hydrochloride (10 mg).
Reaction was allowed to take place by carrying out
reciprocation-shaking at 28.degree. C. and at 300 rpm for 48 hours.
The reaction solution was analyzed on HPLC (Column Sumichiral AGP
manufactured by Sumika Chemical Analysis Service; diameter of 4 mm;
length of 150 mm; 0.03 M sodium phosphate buffer; pH 7.0; flow rate
of 0.5 ml/min; detection wavelength at UV 220 nm). The produced
amounts and the optical activities of pseudoephedrine were
determined. Consequently, only l-pseudoephedrine was obtained
selectively, as shown in Table 6.
TABLE 6 Optical purity (%) Produced Microorganism d- l- d-pseudo-
l-pseudo- amount Example No. genus IFO ephedrine ephedrine
ephedrine ephedrine (mg/ml) Example 49 Corprinus 30197 0 0 0 100
1.09 rhizophorus
Example 50
Production of
(1S,2S)-1-(p-hydroxyphenyl)-2-methylamino-1-propanol
Rhodococcus erythropolis MAK-34 strain was shake-cultured in a
medium (5 ml) containing 1% saccharose, 0.5% corn steep liquor,
0.1% potassium dihydrogenphosphate, 0.3% dipotassium
hydrogenphosphate and 0.1% 1-amino-2-propanol at 30.degree. C. for
48 hours. Either centrifugation or filtration yielded bacterial
cells. To this were added an adequate amount of water, 1M phosphate
buffer (0.2 ml; pH 7.0), glucose (10 mg) and racemic
1-(p-hydroxyphenyl)-2-methylamino-1-propanone hydrochloride (1 mg),
and they were mixed. One milliliter was shaken for reaction at
30.degree. C. for 48 hours. The reaction solution was either
centrifuged or filtered. The supernatant was analyzed on HPLC (.mu.
Bondapakphenyl manufactured by Waters Inc.; diameter of 4 mm;
length of 300 mm; eluent--0.05 M sodium phosphate buffer
(containing 7% acetonitrile); pH 6.5; flow rate of 0.8 ml/min;
detection wavelength at UV 220 nm). Consequently, it was confirmed
that threo-1-(p-hydroxyphenyl)-2-methylamino-1-propanol
hydrochloride was produced at 0.6 mg/ml. To determine the optical
purity of the product, the sample was analyzed on HPLC (Sumichiral
OA-4900 manufactured by Sumika Chemical Analysis Service;
eluent:hexane:dichloroethane:methanol: trifluoroacetic
acid=240:140:40:1; flow rate of 1 ml/min; detection wavelength at
UV 254 nm). Consequently, it was found that
(1S,2S)-1-(p-hydroxyphenyl)-2-methylamino-1-propanol was obtained
in 100% optical purity.
Examples 51 to 54
Production of optically active
1(p-hydroxyphenyl)-2-methylamino-1-propanol
Except that the microorganisms shown in Table 7 were used in place
of Rhodococcus erythropolis MAK-34 strain and the culturing
conditions in the table were followed, reaction was carried out
similarly to Example 50 and optically active
1-(p-hydroxyphenyl)-2-methylamino-1-propanol was obtained. The
results are shown in Table 7. In all instances, the optically
active compound was obtained efficiently.
The culture conditions listed in Tables 7-9 are as follows:
Culture conditions 1: A microorganism was inoculated to a medium
(20 ml) containing 1% glucose, 0.5% peptone, 10 and 0.3% yeast
extract (pH7.0) and culturing was carried out at 30.degree. C. for
48 hours under the shaking condition of 150 rpm.
Culture conditions 2: A microorganism was inoculated to a medium
(20 ml, pH 6.0) containing 5% malt extract and 0.3% yeast extract
and culturing was carried out at 30.degree. C. for 48 hours under
the shaking condition of 150 rpm.
TABLE 7 Produced Stereochemical Example Culture amount
configuration Optical No. Strain IFO conditions (mg/mL) of product
purity (%) Example 51 Helicostylum 8091 1 0.06 1S 2S 90 nigricans
Example 52 Amycolatopsis 14500 1 0.01 1R 2R 100 orientalis lurida
Example 53 Amycolatopsis 12362 1 0.02 1R 2R 100 orientalis
orientalis Example 54 Amycolatopsis 12806 1 0.01 1R 2R 100
orientalis orientalis
Example 55
Production of (1S,2S)-2-ethylamino-1-phenyl-1-propanol
Rhodococcus erythropolis MAK-34 strain was shake-cultured in a
medium (5 ml) containing 1% saccharose, 0.5% corn steep liquor,
0.5% potassium dihydrogenphosphate, 0.3% dipotassium
hydrogenphosphate and 0.1% 1-amino-2-propanol at 30.degree. C. for
48 hours. Either centrifugation or filtration yielded bacterial
cells. To this were added an adequate amount of water, 1M phosphate
buffer (0.2 ml, pH 7.0), glucose (10 mg) and racemic
2-ethylamino-1-phenyl-1-propanone hydrochloride (1 mg), and they
were mixed. One milliliter was shaken for reaction at 30.degree. C.
for 48 hours. The reaction solution was either centrifuged or
filtered. The supernatant was analyzed on HPLC (.mu.
Bondaspherephenyl manufactured by Waters Inc.; diameter of 4 mm;
length of 150 mm; eluent--0.05 M sodium phosphate buffer
(containing 7% acetonitrile); pH 6.5; flow rate of 0.8 ml/min;
detection wavelength at UV 220 nm). Consequently, it was confirmed
that threo-2-ethylamino-1-phenyl-1-propanol hydrochloride was
produced at 0.47 mg/ml. To determine the optical purity of the
product, the sample was analyzed on HPLC (Column OD manufactured by
Daicel Chemical Industries Ltd.; diameter of 4.6 mm; length of 250
mm; eluent-hexane:isopropanol:diethylamine=90:10:0.1; flow rate of
1 ml/min; detection wavelength at UV 254 nm). Consequently, the
product was found to be (1S,2S)-2-ethylamino-1-phenyl-1-propanol
(optical purity: 100%).
Examples 56 to 58
Production of (1R,2R)-2-ethylamino-1-phenyl-1-propanol
Except that the microorganisms shown in Table 8 were used in place
of Rhodococcus erythropolis MAK-34 strain and the culturing
conditions in the table were followed, reaction was carried out
similarly to Example 55 and
(1R,2R)-2-ethylamino-1-phenyl-1-propanol was obtained. The results
are shown in Table 8.
TABLE 8 Produced Stereochemical Example Culture amount
configuration Optical No. Strain IFO conditions (mg/mL) of product
purity (%) Example 56 Amycolatopsis 15602 1 0.01 1R 2R 100 alba
Example 57 Amycolatopsis 12806 1 0.01 1R 2R 100 orientalis
orientalis Example 58 Rhodotorula 0951 2 0.01 1R 2R 100
aurantiaca
Examples 59 to 61
Production of Optically Active
1-(m-hydroxyphenyl)-2-amino-1-propanol
The microorganisms listed in Table 9 were shake-cultured in a
medium (5 ml) at 30.degree. C. for 48 hours under their respective
conditions. Either centrifugation or filtration yielded bacterial
cells. To this were added an adequate amount of water, 1M phosphate
buffer (0.2 ml, pH 7.0), glucose (10 mg), and racemic
1-(m-hydroxyphenyl)-2-amino-1-propanone hydrochloride (1 mg), and
they were mixed. One milliliter was shaken for reaction at
30.degree. C. for 48 hours. This was either centrifuged or
filtered. The supernatant was analyzed on HPLC (.mu. Bondapakphenyl
manufactured by Waters Inc.; diameter of 4 mm; length of 300 mm;
eluent--0.05 M sodium phosphate buffer (containing 7%
acetonitrile); pH 6.5; flow rate of 0.8 ml/min; detection
wavelength at UV 220 nm). Consequently, it was confirmed that
threo-1-(m-hydroxyphenyl)-2-amino-1-propanol was produced. To
determine the optical purity of the product, the sample was
analyzed on HPLC (Crownpak CR+ manufactured by Daicel Chemical
Industries Ltd.; perchloric acid, pH 2.0, 1.0 ml/min, UV 254 nm).
Consequently, it was found that optical active
1-(m-hydroxyphenyl)-2-amino-1-propanol with its stereochemical
configuration shown in Table 9.
TABLE 9 Produced Stereochemical Optical Example Culture amount
configuration of purity No. Strain IFO conditions (mg/mL) product
(%) Example 59 Helicostylums 8091 1 0.01 1S 2S 100 nigricans
Example 60 Amycolatopsis 15602 1 0.01 1R 2R 78 albas Example 61
Rhodotorula 0951 2 0.01 1R 2R 100 aurantiaca
Example 62
Production of (1R,2R)-1-(p-hydroxyphenyl)-2-amino-1-propanol
Amycolatopsis alba IFO-15602 was cultured under culture conditions
1, and either centrifugation or filtration yielded bacterial cells.
To these were added an adequate amount of water, 1M phosphate
buffer (0.2 ml, pH 7.0), glucose (10 mg) and racemic
1-(p-hydroxyphenyl)-2-amino-1-propanone hydrochloride (1 mg), and
they were mixed. One milliliter was shaken for reaction at
30.degree. C. for 48 hours. This was either centrifuged or
filtered. The supernatant was analyzed on HPLC (.mu. Bondapakphenyl
manufactured by Waters Inc.; diameter of 4 mm; length of 300 mm;
eluent-0.05 M sodium phosphate buffer (containing 7% acetonitrile);
pH 6.5; flow rate of 0.8 ml/min; detection wavelength at UV 220
nm). Consequently, it was confirmed that
threo-1-(p-hydroxyphenyl)-2-amino-1-propanol hydrochloride was
produced at 0.03 mg/ml. To determine the optical purity of the
product, the sample was analyzed on HPLC (Crownpak CR+ manufactured
by Daicel Chemical Industries Ltd.; perchloric acid, pH 2.0, 1.0
ml/min, UV 254 nm). Consequently, the product was found to be
(1R,2R)-1-(p-hydroxyphenyl)-2-amino-1-propanol (optical purity:
82%).
Example 63
Production of (1S,2S)-1-phenyl-2-amino-1-butanol
Helicostylum nigricans IFO-8091 was cultured under culture
conditions 1. Either centrifugation or filtration yielded bacterial
cells. To these were added an adequate amount of water, 1M
phosphate buffer (0.2 ml, pH 7.0), glucose (10 mg) and racemic
1-phenyl-2-amino-1-butanone hydrochloride (1 mg), and they were
mixed. One milliliter was shaken for reaction at 30.degree. C. for
48 hours. The reaction solution was either centrifuged or filtered.
The supernatant was analyzed on HPLC (.mu. Bondaspherephenyl
manufactured by Waters Inc.; diameter of 4 mm; length of 150 mm;
eluent-0.05 M sodium phosphate buffer (containing 7% acetonitrile);
pH 6.5; flow rate of 0.8 ml/min; detection wavelength at UV 220
nm). Consequently, it was found that
threo-1-phenyl-2-amino-1-butanol hydrochloride was produced at 0.62
mg/ml. To determine the optical purity of product, the sample was
analyzed on HPLC (OD by Daicel Chemical Industries Ltd.; diameter
of 4.6 mm; length of 250 mm;
hexane:isopropanol:diethylamine=90:10:0.1; 1 ml/min; UV 254 nm).
Consequently, the product was found to be
(1S,2S)-1-phenyl-2-amino-1-butanol.
Example 64
Production of (1R,2R)-1-phenyl-2-amino-1butanol
Amycolatopsis orientalis IFO-12806 was cultured under culture
conditions 1, and similarly to Example 63,
(1R,2R)-1-phenyl-2-amino-1-butanol could be produced at 0.21
mg/ml.
Example 65
The Effect of Addition of Inducer (1)
1-Amino-2-hydroxypropanone was added to medium 1 (Table 10) so that
a level of 5 g/L could be obtained. Five milliliters was then
poured into a test tube. With a silicone stopper it was sterilized
in an autoclave at 121.degree. C. for 30 minutes. The
microorganisms listed in Table 11 were inoculated to this medium
and to the medium with no addition of the inducer, respectively;
and they were shake-cultured at 300 rpm and at 30.degree. C. for 48
hours. The culture (0.5 mL) was centrifuged at 10,000 G for 20
minutes. The bacterial cells obtained by removal of the supernatant
were suspended by addition of water to prepare a uniform
suspension. To this were added water, buffer and
dl-2-methylamino-1-phenyl-1-propanone hydrochloride (10 mg),
forming 1 mL. The one milliliter was poured into a test tube and
reaction was allowed to take place under shaking at 150 rpm and at
30.degree. C. for 12 hours. After the reaction, the bacterial cells
were removed by centrifugation and the supernatant was subjected to
HPLC, whereby the produced amount of pseudoephedrine was determined
(HPLC conditions: .mu. Bondapakphenyl manufactured by Waters Inc.;
diameter of 4 mm; length of 300 mm; eluent--0.05 M sodium phosphate
buffer (containing 7% acetonitrile); pH 6.5; flow rate of 0.8
ml/min; detection wavelength at UV 220 nm).
As the results are shown in Table 11, the produced amounts of
pseudoephedrine when culturing was carried out with the addition of
the inducer displayed remarkable increases as compared to the
culturing with no addition of inducer.
TABLE 10 Composition of Composition of Composition of Composition
of medium 1 medium 2 medium 3 medium 4 saccharose 1% glucose 0.1%
glucose 1% soluble starch 1% corn steep tryptone 0.5% Bactopeptone
glucose 0.5% liquor 0.5% 0.5% potassium yeast yeast Nzaminetype
dihydrogen- extract 0.5% extract 0.3% A 0.3% phosphate 0.1%
dipotassium dipotassium pH 7.0 tryptone 0.5% hydrogen- hydrogen-
phosphate 0.3% phosphate p-aminobenzoic pH 7.0 yeast acid 0.01%
extract 0.2% H 7.0 dipotassium hydrogen- phosphate 0.1% magnesium
sulfate 7H.sub.2 O 0.05%
TABLE 11 Produced amount Produced (no amount addition) (addition)
No. Microorganism No. Medium mg mg 1 Rhodococcus MAK-34 1 0.018
1.26 erythropolis 2 Mycobacterium IFO-15527 3 0.032 0.77
chlorophenolicum 3 Mycobacterium IFO-12065 3 0.048 0.21 smegmatis 4
Nocardioides IFO-12069 2 0 0.19 simplex 5 Klebsiella IFO-3319 2
0.018 0.066 pneumoniae 6 Absidia IFO-4409 4 0.0035 0.22 lichtheimi
7 Aspergillus IFO-4033 4 0.00048 1.17 awamori 8 Aspergillus
IFO-5468 4 0.0092 0.018 candidus 9 Penicillium IFO-5337 4 0.031
1.26 cyaneum 10 Hypocrea IFO-9165 4 0.0058 0.64 gelatinosa 11
Helicostylum IFO-8091 4 0.0067 0.52 nigricans 12 Tritirachium
IFO-7544 4 0.0047 0.078 oryzae 13 Armillariella IFO-31616 4 0.0042
0.46 mellea
Example 66
The Effect of Addition of Inducers (2)
Rhodococcus erythropolis MAK-34 was inoculated to 5 ml of a medium
(pH 7.0) containing 1.0% saccharose, 0.5% corn steep liquor, 0.1%
dipotassium hydrogenphosphate, 0.3% potassium dihydrogenphosphate,
0.01% p-aminobenzoic acid and each inducer; and shake-culturing was
carried out at 30.degree. C. for 48 hours. After the culture was
centrifuged to give bacterial cells, they were placed into a test
tube and suspended by adding 1.0 ml of 0.2 M sodium phosphate
buffer (pH 7.0) thereto. To this were added
dl-2-methylamino-1-phenyl-1-propanone hydrochloride (10 mg) and
glucose (20 mg) and reaction was allowed to take place under
shaking at 30.degree. C. for 16 hours. After the reaction, the
reaction solution was centrifuged to remove bacterial cells, and
the supernatant was subjected to HPLC, producing optically active
pseudoephedrine (.mu. Bondaspherephenyl manufactured by Waters
Inc.; diameter of 4 mm; length of 150 mm; eluent--7%
acetonitrile-0.05 M sodium phosphate buffer (pH 6.5); flow rate of
0.8 ml/min; detection wavelength at 220 nm). As shown in Table 12,
the production displayed remarkably higher values than does the
case without the addition of inducer.
TABLE 12 Compound name Produced amount (mg)
1-acetylamino-2-propanol 3.00 1-methylamino-2-propanol 2.83
1-amino-2-oxopropane 1.97 2-amino-3-hydroxybutane 0.05
1-amino-2-hydroxybutane 0.65 no addition 0.02
Comparative Example 1
Except that Brettanomyces anomalus IFO 0642 was used instead of
Microbacterium arborescens IFO 3750, the production reaction of
pseudoephedrine was attempted similarly to Example 1. However, no
reduced product was obtained.
Comparative Example 2
Except that Candida guilliermondii IFO 0566 was used instead of
Microbacterium arborescens IFO 3750, the production reaction of
pseudoephedrine was attempted similarly to Example 1. However, no
reduced product was obtained.
Comparative Example 3
Except that Schizosaccharomyces pombe IFO 0358 was used instead of
Microbacterium arborescens IFO 3750, the production reaction of
pseudoephedrine was attempted similarly to Example 1. However, no
reduced product was obtained.
Comparative Example 4
Except that Bacillus subtilis IFO 3037 was used instead of
Microbacterium arborescens IFO 3750, the production reaction of
pseudoephedrine was attempted similarly to Example 1. However, no
reduced product was obtained.
Industrial Applicability
As described above, the process for producing an optically active
.beta.-amino alcohol according to this invention allows the
.beta.-amino alcohol having the desired optical activity to be
produced from an enantiomeric mixture of an .alpha.-aminoketone
compound or a salt thereof in a high yield as well as in a highly
selective manner with a simple process while sufficiently
preventing the generation of diastereomeric byproducts.
Accordingly, this invention will make it possible to produce
pseudoephedrines, among others, having the desired optical activity
in a high yield as well as in a highly selective manner and thus it
is valuable in the manufacture of drugs and their
intermediates.
* * * * *